We have being using these columns for a couple of months. Tend to re-use them according to salt (if it was used to desalt for DTT, for instance) and protein used. We do wash them with ddH2O and then store at RT in 20%ethanol. But I'm not sure how many more times we can do this and still get a good performance.
Anyone has had experience with this ?
Typically, I use theme once. However, if you want to re-use them you need to make sure that their capacity does not decline. For example, apply a solution containing known amount of protein and check how much is recovered. For desalting capacity, switch to a tris buffer and check the pH
Lots, if stored carefully. Check from time to time using a coloured molecule that there is no formation of preferential pathways (often occurs if you let them dry). I often add azide for storage.
Dear Inna,
My experience has also been limited to one time usage and find them very efficient. Cytochrome C which is a red colored protein and it is what we trypically use to see the performance of columns that are used for polypeptide separations. It is from horse and very cheap to purchase. You can make up stock solutions (eg;, 5 microgrammes per microliters) and use a couple of microliters of this solution on your test column. You should be able to follow the red circle of protein as it is migrating over the column packing. It will tell you how the column resolution and binding/elution efficiency of the column is. If you do not want to worry about carryover, just use Cytochrome C to fine tune and check column performance once.
They come stored in azide so as Catherine mentioned, having azide in your storage solution will help.
Good luck,
Hediye.
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